Quinolones have been shown to be effective to varying degrees against a range of bacterial pathogens. However, as diseases caused by these pathogens are on the rise, there exists a need for antimicrobial compounds that are more potent than the present group of quinolones.
Gemifloxacin mesylate (SB-265805) is a novel fluoroquinolone useful as a potent antibacterial agent. Gemifloxacin compounds are described in detail in patent application PCT/KR98/00051 published as WO 98/42705. Patent application EP 688772 discloses novel quinolone(naphthyridine)carboxylic acid derivatives, including anhydrous (R,S)-7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid of formula I. 
PCT/KR98/00051 discloses (R,S)-7-(3-aminomethyl-4-syn-methoxyimino-pyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid methanesulfonate and hydrates thereof including the sesquihydrate.
I. Pneumococcal Pathogens
The incidence of pneumococci resistant to penicillin G and other xcex2-lactam and non-xcex2-lactam compounds has increased worldwide at an alarming rate, including in the U.S. Major foci of infections currently include South Africa, Spain, Central and Eastern Europe, and parts of Asia (P. C. Appelbaum, Clin. Infect. Dis. 15:77-83, 1992; Friedland, et al., Pediatr. Infect. Dis. 11:433-435, 1992; Friedland, et al., N. Engl. J. Med. 331:377-382, 1994; Jacobs, et al., Clin. Infect. Dis. 15:119-127, 1992 and Jacobs, et al., Rev. Med. Microbiol. 6:77-93, 1995). In the U.S. a recent survey has shown an increase in resistance to penicillin from  less than 5% before 1989 (including  less than 0.02% of isolates with MICs xe2x89xa72.0 xcexcg/ml) to 6.6% in 1991-1992 (with 1.3% of isolates with MICs xe2x89xa72.0 xcexcg/ml) (Brieman, et al., J. Am. Med. Assoc. 271:1831-1835, 1994). In another more recent survey, 23.6% (360) of 1527 clinically significant pneumococcal isolates were not susceptible to penicillin (Doern, et al., Antimicrob. Agents Chemother. 40:1208-1213, 1996). It is also important to note the high rates of isolation of penicillin intermediate and resistant pneumococci (approximately 30%) in middle ear fluids from patients with refractory otitis media, compared to other isolation sites (Block, et al., Pediatr. Infect. Dis. 14:751-759, 1995). The problem of drug-resistant pneumococci is compounded by the ability of resistant clones to spread from country to country, and from continent to continent (McDougal, et al., Antimicrob. Agents Chemother. 36:2176-2184, 1992; Munoz, et al., Clin. Infect. Dis. 15:112-118, 1992).
There is an urgent need of oral compounds for out-patient treatment of otitis media and respiratory tract infections caused by penicillin intermediate and resistant pneumococci (Friedland, et al., Pediatr. Infect. Dis. 11:433-435, 1992; Friedland, et al., N. Engl. J. Med. 331:377-382, 1994; M. R. Jacobs, Clin. Infect. Dis. 15:119-127, 1992; and Jacobs, et al., Rev. Med. Microbiol. 6:77-93, 1995). Available quinolones such as ciprofloxacin and ofloxacin yield moderate in vitro activity against pneumococci, with MICs clustering around the breakpoints (Spangler, et al., Antimicrob. Agents Chemother. 36:856-859, 1992; and Spangler, et al., J. Antimicrob. Chemother. 31:273-280, 1993). Gemifloxacin (SB 265805)(LB 20304a) is a new broad-spectrum fluoronaphthyridone carboxylic acid with a novel pyrrolidone substituent (Cormican, et al., Antimicrob. Agents Chemother. 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; and Oh, et al., Antimicrob. Agents Chemother. 40:1564-1568, 1996). Previous preliminary studies (Cormican, et al., Antimicrob. Agents Chemother. 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; and Oh, et al., Antimicrob. Agents Chemother. 40:1564-1568, 1996) have shown that this compound is very active against pneumococci. This study further examined the antipneumococcal activity of gemifloxacin compared to ciprofloxacin, levofloxacin, sparfloxacin, grepafloxacin, trovafloxacin, amoxicillin, cefuroxime, azithromycin and clarithromycin by i) agar dilution testing of 234 quinolone susceptible and resistant strains; ii) examination of resistance mechanisms in quinolone resistant strains; iii) time-kill testing of 12 strains; iv) examination of the post-antibiotic effect (herein xe2x80x9cPAExe2x80x9d) of drugs against 6 strains.
Provided herein is a significant discovery made using a gemifloxacin compound against a range of penicillin susceptible and resistant pneumococci by agar dilution, microdilution, time-kill and post-antibiotic effect methodology. Against 64 penicillin susceptible, 68 intermediate and 75 resistant pneumococci (all quinolone susceptible), agar dilution MIC50/90 values (xcexcg/ml) were as follows: gemifloxacin, 0.03/0.06; ciprofloxacin, 1.0/4.0; levofloxacin, 1.0/2.0; sparfloxacin, 0.5/0.5; grepafloxacin, 0.125/0.5; trovafloxacin, 0.125/0.25; amoxicillin, 0.016/0.06 (penicillin susceptible), 0.125/1.0 (penicillin intermediate), 2.0/4.0 (penicillin resistant); cefuroxime, 0.03/0.25 (penicillin susceptible), 0.5/2.0 (penicillin intermediate), 8.0/16.0 (penicillin resistant); azithromycin, 0.125/0.5 (penicillin susceptible), 0.125/ greater than 128.0 (penicillin intermediate), 4.0/ greater than 128.0 (penicillin resistant); clarithromycin, 0.03/0.06 (penicillin susceptible), 0.03/32.0 (penicillin intermediate), 2.0/ greater than 128.0 (penicillin resistant). Against 28 strains with ciprofloxacin MICs xe2x89xa78 xcexcg/ml, gemifloxacin had the lowest MICs (0.03-1.0 xcexcg/ml, MIC90 0.5 xcexcg/ml), compared with MICs ranging between 0.25 to  greater than 32.0 xcexcg/ml)(MIC90s 4.0xe2x86x9232.0 xcexcg/ml) for the other quinolones. Resistance in these 28 strains was associated with mutations in parC, gyrA, parE, and/or gyrb or efflux, with some strains having multiple resistance mechanisms. For 12 penicillin susceptible and resistant pneumococcal strains (2 quinolone resistant), time-kill results showed that levofloxacin at the MIC, gemifloxacin and sparfloxacin at 2xc3x97MIC and ciprofloxacin, grepafloxacin and trovafloxacin at 4xc3x97MIC, were bactericidal after 24 h. Gemifloxacin was uniformly bactericidal after 24 h at xe2x89xa60.5 xcexcg/ml. Various degrees of 90% and 99% killing by all quinolones was detected after 3 h. Gemifloxacin and trovafloxacin were both bactericidal at the microbroth MIC for the two quinolone resistant pneumococcal strains. Amoxicillin, at 2xc3x97MIC and cefuroxime at 4xc3x97MIC, were bactericidal after 24 h, with some degree of killing at earlier time periods. By contrast, macrolides gave slower killing against the 7 susceptible strains tested, with 99.9% killing of all strains at 2-4xc3x97MIC after 24 h. Post-antibiotic effects for 5 quinolone susceptible strains were similar (0.3-3.0 h) for all quinolones tested, and significant quinolone PAEs were found for the quinolone-resistant strain.
Also provided herein is a significant discovery made using a gemifloxacin compound against quinolone-resistant pneumococci, demonstrating the activity of the gemifloxacin compound used was superior to a number of quinolones as described in more detail herein. Gemifloxacin compounds are valuable compounds for the treatment of infections caused by a range of pneumococcal pathogens, including those resistant to usual oral therapy, thereby filling an unmet medical need.
II. Haemophilus Pathogens
Although development of an effective vaccine against Haemophilus influenzae type b has led to disappearance of this organism in many parts of the world, its place has been taken by untypeable H. influenzae strains. The latter organisms (followed by Streptococcus pneumoniae and Moraxella catarrhalis) are now considered to be the leading cause of acute exacerbations of chronic bronchitis, and an important cause, together with S. pneumoniae and M. catarrhalis, of acute otitis media, sinusitis and conmmunity-acquired respiratory tract infections (Fang, et al., Medicine (Baltimore) 69:307-316, 1990; Hoberman, et al., Pediatr. Infect. Dis. 10:955-962, 1996; Jacobs, et al., Antimicrob. Agents Chemother, In press; and Zeckel, et al., Clin. Ther. 14:214-229, 1992).
Current recommendations by the NCCLS for use of Haemophilus Test Medium (herein xe2x80x9cHTMxe2x80x9d) for Haemophilus susceptibility testing have been complicated by difficulty in commercial manufacture of this medium, and its short balf-life when made in-house. Reliable Haemophilus susceptibility testing with HEM requires use of freshly made medium used within 3 weeks of making (Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, 3rd Edition, NCCLS, Wayne, Pa., 1997).
Previous preliminary studies have shown that this gemifloxaxin is very active against Haemophilus and Moraxella (Cormican, et al., Antimicrob. Agents Chemother. 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; and Oh, et al., Antimicrobial Agents Chemother. 40:1564-1568, 1996).
A further embodiment provided herein is based in part on a significant discovery made using a gemifloxacin compound against nine rare clinical strains of Haemophilus influenzae from Europe with increased ciprofloxacin MICs were tested for in vitro activity (MICs) of gemifloxacin (SB-265805), ciprofloxacin, levofloxacin, sparfloxacin, grepafloxacin and trovafloxacin and checked for mutations in gyrA, parC, gyrB and parE, demonstrating the activity of the gemifloxacin compound used was superior to a number of quinolones as described in more detail herein. Gemifloxacin compounds are valuable compounds for the treatment of infections caused by a range of Haemophilus influenzae strains, including those resistant to usual oral therapy, thereby filling an unmet medical need.
I. Pneumococcal Pathogens
An object of the invention is a method for modulating metabolism of pneumococcal pathogenic bacteria comprising the step of contacting pneumococcal pathogenic bacteria with an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound, or an antibacterially effective derivative thereof.
A further object of the invention is a method wherein said pneumococcal pathogenic bacteria is selected from the group consisting of: bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrb; bacteria comprising a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; bacteria comprising a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; bacteria comprising a mutation in parE at D435-N or I460-V; bacteria comprising a mutation in gyrB at D435-N or E474-K; bacteria comprising at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in parE at D435-N or I460-V; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in gyrB at D435-N or E474-K; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB; Streptococcus pneumoniae bacteria comprising a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; Streptococcus pneumoniae bacteria comprising a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116G; Streptococcus pneumoniae bacteria comprising a mutation in pare at D435-N or I460-V; Streptococcus pneumoniae bacteria comprising a mutation in gyrB at D435-N or E474-K; Streptococcus pneumoniae bacteria comprising at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; and Streptococcus pneumoniae bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB.
Also provided by the invention is a method of treating or preventing a bacterial infection by pneumococcal pathogenic bacteria comprising the step of administering an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound to a mammal suspected of having or being at risk of having an infection with pneumococcal pathogenic bacteria.
A preferred method is provided wherein said modulating metabolism is inhibiting growth of said bacteria or killing said bacteria.
A further preferred method is provided wherein said contacting said bacteria comprises the further step of introducing said composition into a mammal, particularly a human.
Further preferred methods are provided by the invention wherein said bacteria is selected from the group consisting of: bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB; bacteria comprising a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; bacteria comprising a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; bacteria comprising a mutation in parE at D435-N or I460-V; bacteria comprising a mutation in gyrB at D435-N or E474-K; bacteria comprising at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in pare at D435-N or I460-V; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in gyrB at D435-N or E474-K; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in a quinolone resistance determining region (QRDR) of parC, gyrA, parE, and/or gyrB; Streptococcus pneumoniae bacteria comprising a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; Streptococcus pneumoniae bacteria comprising a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; Streptococcus pneumoniae bacteria comprising a mutation in parE at D435-N or I460-V; Streptococcus pneumoniae bacteria comprising a mutation in gyrB at D435-N or E474-K; Streptococcus pneumoniae bacteria comprising at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; and Streptococcus pneumoniae bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parc, gyrA, parE, and/or gyrB.
Also provided is a method for modulating the activity of a topoisomerase comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA or parE or gyrB.
It is preferred in the methods of the invention that said mutation in ParC is at S79-F or Y, D83-N, R95-C, or K137-N; said mutation in gyrA is at S83-A, C, F, or Y; E87-K; or S116-G; said mutation in parE is at D435-N or I460-V; or said mutation in gyrB is at D435-N or E474-K.
An object of the invention is a method for modulating metabolism of quinolone-resistant pneumococcal pathogenic bacteria comprising the step of contacting quinolone-resistant pneumococcal pathogenic bacteria with an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound, or an antibacterially effective derivative thereof.
A further object of the invention is a method wherein said quinolone-resistant pneumococcal pathogenic bacteria is selected from the group consisting of: a pneumococcal strain comprising a mutation in the quinolone resistance-determining region (QRDR) of parC and/or gyrA; a pneumococcal strain comprising a mutation in parC said mutation comprising S79-F and/or Y, D83-G and/or N, N91-D, R95-C, and/or K137-N; a pneumococcal strain comprising a mutation in gyrA said mutation comprising S81-A, C, F, or Y; E85-K; and/or S114-G; a pneumococcal strain comprising a mutation in parE said mutation comprising D43 5-N and/or I460-V; a pneumococcal strain comprising a mutation in gyrB said mutation comprising D435-N and/or E474-K; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA,parE, and/or gyrB; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA,parE, and/or gyrB, any of which are resistant to ciprofloxacin, levofloxacin, or sparfloxacin; and a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB, any of which also comprising an efflux mechanism of quinolone resistance.
Also provided by the invention is a method of treating or preventing a bacterial infection by quinolone-resistant pneumococcal pathogenic bacteria comprising the step of administering an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound to a mammal suspected of having or being at risk of having an infection with quinolone-resistant pneumococcal pathogenic bacteria.
Further preferred methods are provided by the invention wherein said bacteria is selected from the group consisting of: a pneumococcal strain comprising a mutation in the quinolone resistance-determining region (QRDR) of parC and/or gyrA; a pneumococcal strain comprising a mutation in parC said mutation comprising S79-F and/or Y, D83-G and/or N, N91-D, R95-C, and/or K137-N; a pneumococcal strain comprising a mutation in gyrA said mutation comprising S81-A, C, F, and/or Y; E85-K; and/or S114-G; a pneumococcal strain comprising a mutation in parE said mutation comprising D435-N and/or I460-V; a pneumococcal strain comprising a mutation in gyrB said mutation comprising D435-N and/or E474-K; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB, any of which are resistant to ciprofloxacin, levofloxacin, or sparfloxacin; and a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB, any of which also comprising an efflux mechanism of quinolone resistance.
II. Haemophilus Pathogens
An object of the invention is a method for modulating metabolism of a rare Haemophilus influenzae strain comprising the step of contacting a rare Haemophilus influenzae strain with an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound, or an antibacterially effective derivative thereof.
A further object of the invention is a method wherein said rare pathogenic H. influenzae strain is selected from the group consisting of: bacteria comprising a mutation set forth in Table 11 or 12; a Haemophilus influenzae strain set forth in Table 11 or 12; bacteria of the genus Haemophilus comprising a mutation set forth in Table 11 or 12; and bacteria of the species Haemophilus influenzae comprising a mutation set forth in Table 11 or 12.
Also provided by the invention is a method of treating or preventing a bacterial infection by a rare pathogenic H. influenzae strain comprising the step of administering an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound to a mammal suspected of having or being at risk of having an infection with a rare pathogenic H. influenzae strain.
A preferred method is provided wherein said modulating metabolism is inhibiting growth of said bacteria or killing said bacteria.
A further preferred method is provided wherein said contacting said bacteria comprises the further step of introducing said composition into a mammal, particularly a human.
Further preferred methods are provided by the invention wherein said bacteria is selected from the group consisting of: bacteria comprising a mutation set forth in Table 11 or 12; a Haemophilus influenzae strain set forth in Table 11 or 12; bacteria of the genus Haemophilus comprising a mutation set forth in Table 11 or 12; and bacteria of the species Haemophilus influenzae comprising a mutation set forth in Table 11 or 12.
Various changes and modifications within the spirit and scope of the disclosed invention will become readily apparent to those skilled in the art from reading the following descriptions and from reading the other parts of the present disclosure.
I. Pneumococcal Pathogens
The present invention provides, among other things, methods for using a composition comprising a quinolone, particularly a gemifloxacin compound against a number of pathogenic bacteria including, for example, strains of Streptococcus pneumoniae and Haemophilus influenzae. 
The present invention firther provides methods for using a composition comprising a quinolone, particularly a gemifloxacin compound against a quinolone-resistant pneumococcal strain, particularly a strain comprising a mutation in the quinolone resistance-determining region (QRDR) of parC and/or gyrA; a pneumococcal strain comprising a mutation in parC said mutation comprising S79-F and/or Y, D83-G and/or N, N91-D, R95-C, and/or K137-N; a pneumococcal strain comprising a mutation in gyrA said mutation comprising S81-A, C, F, and/or Y; E85-K; and/or S114-G; a pneumococcal strain comprising a mutation in parE said mutation comprising D435-N and/or I460-V; a pneumococcal strain comprising a mutation in grB said mutation comprising D435-N and/or E474-K; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA,parE, and/or gyrB, any of which are resistant to ciprofloxacin, levofloxacin, or sparfloxacin; and a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gerA, parE, and/or gyrB, any of which also comprising an efflux mechanism of quinolone resistance.
As used herein xe2x80x9cgemifloxacin compound(s)xe2x80x9d means a compound having antibacterial activity described in patent application PCT/KR98/00051 published as WO 98/42705, or patent application EP 688772.
Previous studies have shown gemifloxacin to be 32 to 64 fold more active than ciprofloxacin, ofloxacin, sparfloxacin and trovafloxacin against methicillin-susceptible and -resistant Staphylococcus aureus, methicillin-resistant Staphylococcus epideridis and S. pneumoniae. Gemifloxacin was also highly active against most members of the family Enterobacteriaceae, with activity was more potent than those of sparfloxacin and ofloxacin and comparable to that of ciprofloxacin. Gemifloxacin was the most active agent against Gram-positive species resistant to other quinolones and glycopeptides. Gemifloxacin has limited activity against anaerobes (Cormican, et al., Antimicrob. Agents Chemother. 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; Oh, et al., Antimicrob. Agents Chemother. 40:1564-1568, 1996).
This invention was based, in part, on analyses evaluating the comparative activity of gemifloxacin against various pneumococcal pathogens. In these analyses, gemifloxacin gave the lowest quinolone MICs against all pneumococcal strains tested followed by trovafloxacin, grepafloxacin, sparfloxacin, levofloxacin and ciprofloxacin. MICs were similar to those described previously (Cormican, et al., Antimicrob. Agents Chemother. 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; Oh, et al., Antimicrob. Agents Chemother. 40:1564-1568, 1996). Additionally, gemifloxacin gave significantly lower MICs against highly quinolone resistant pneumococci, irrespective of quinolone resistance mechanism. This was the case in double mutants with mutations in both parC and gyrA, strains which have previously been shown to be highly resistant to other quinolones, as well as for strains with an efflux mechanism (Brenwald, et al., Antimicrob. Agents Chemother. 42:2032-2035, 1998; and Pan et al., Antimicrob. Agents Chemother. 40:2321-2326, 1996). MICs of non-quinolone agents were similar to those described previously (M. R. Jacobs, Clin. Infect. Dis. 15:119-127, 1992; Jacobs, et al., Rev. Med. Microbiol. 6:77-93, 1995; Pankuch, et al., J. Antimicrob. Chemother. 35:883-888, 1995).
Gemifloxacin also showed good killing against the 12 strains tested, including the two quinolone resistant strains. At xe2x89xa60.5 xcexcg/ml, gemifloxacin was bactericidal against all 12 strains. Killing rates relative to MICs were similar to those of other quinolones, with significant killing occurring earlier than xcex2-lactams and macrolides. Kill kinetics of quinolone and non-quinolone compounds in the analyses described herein were similar to those described previously (Pankuch, et al., Antimicrob. Agents Chemother. 38:2065-2072, 1994; Pankuch et al., Antimicrob. Agents Chemother. 40:1653-1656, 1996; and Visalli, et al., Antimicrob. Agents Chemother. 40:362-366, 1996). Gemifloxacin also gave, together with the other quinolones tested, significant PAEs against all 6 strains tested, including the one quinolone resistant strain. The higher ciprofloxacin PAE at both exposure concentrations is of no significance, because, with an MIC of 32 xcexcg/ml, 5xc3x97 and 10xc3x97MIC are not clinically achievable with this strain. PAE values for quinolones and macrolides were similar to those described previously (Fuursted, et al., Antimicrob. Agents Chemother. 41:781-784, 1997; Licata, et al., Antimicrob. Agents Chemother. 41:950-955, 1997; Spangler, et al., Antimicrob. Agents Chemother. 41:2173-2176, 1997; and Spangler, et al., Antimicrob. Agents Chemother. 42:1253-1255, 1998). It is generally accepted that quinolones have similar PAEs against pneumococci.
In summary, gemifloxacin was the most potent quinolone tested by MIC and time-kill against both quinolone susceptible and resistant pneumococci and, similar to other quinolones, gave PAEs against quinolone susceptible strains. The incidence of quinolone resistant pneumococci is currently very low. However, this situation may change with the introduction of broad-spectrum quinolones into clinical practice, and in particular in the pediatric population, leading to selection of quinolone resistant strains (Davies, et al., Antimicrob. Agents Chemother. 43:1177-1182, 1999). Gemifloxacin is a promising new antipneumococcal agent against pneumococci, irrespective of their susceptibility to quinolones and other agents. Clinical studies will be necessary in order to validate this hypothesis.
Results of agar dilution MIC testing of the 207 strains with ciprofloxacin MICs xe2x89xa64.0 xcexcg/ml are presented in Table 1. MIC50/90 values (xcexcg/ml) were as follows: gemifloxacin, 0.03/0.06; ciprofloxacin, 1.0/4.0; levofloxacin, 1.0/2.0; sparfloxacin, 0.5/0.5; grepafloxacin, 0.125/0.5; trovafloxacin, 0.125/0.25; amoxicillin, 0.016/0.06 (penicillin susceptible), 0.125/1.0 (penicillin intermediate), 2.0/4.0 (penicillin resistant); cefuroxime, 0.03/0.25 (penicillin susceptible), 0.5/2.0 (penicillin intermediate), 8.0/16.0 (penicillin resistant); azithromycin, 0.125/0.5 (penicillin susceptible), 0.125/ greater than 128.0 (penicillin intermediate), 4.0/ greater than 128.0 (penicillin resistant); clarithromycin, 0.03/0.06 (penicillin susceptible), 0.03/32.0 (penicillin intermediate), 2.0/ greater than 128.0 (penicillin resistant). Against 28 strains with ciprofloxacin MICs xe2x89xa78 xcexcg/ml, gemifloxacin had the lowest MICs (0.03-1.0 xcexcg/ml, MIC90 0.5 xcexcg/ml), compared with MICs ranging between 0.25 to  greater than 32.0 xcexcg/ml)(MIC90s 4.0xe2x86x9232.0 xcexcg/ml) for the other quinolones, with trovafloxacin, grepafloxacin, sparfloxacin and levofloxacin, in ascending order, giving the next lowest MICs (Table 2). Mechanisms of quinolone resistance are presented in Tables 3 and 4. As can be seen, quinolone resistance was associated with mutations in the quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB. Mutations in ParC were at S79-F or Y, D83-N, R95-C, or K137-N. Mutations in gyrA were at S83-A, C, F, or Y; E87-K; or S116-G. Twenty one strains had a mutation in parE at D435-N or I460-V. Only two strains had a mutation in gyrB at D435-N or E474-K. Twenty strains had a total of three or four mutations in the QRDRs or parC, gyrA, parE, and gyrB (Table 3). Amongst these 20 strains all were resistant to ciprofloxacin (MICs  greater than 8 xcexcg/ml), levofloxacin (MICs  greater than 4 xcexcg/ml), and sparfloxacin (MICs  greater than 1 xcexcg/ml); 19 were resistant to grepafloxacin (MICs  greater than 1 xcexcg/ml); and 10 were resistant to trovafloxacin (MICs  greater than 2 xcexcg/ml), yet gemifloxacin MICs were  less than 0.5 xcexcg/ml in 18 of the strains (Table 2).
In the presence of reserpine 23 strains had lower ciprofloxacin MICs (2-16xc3x97); 13 strains had lower gemifloxacin MICs (2-4xc3x97); 7 strains had lower levofloxacin MICs (2-4xc3x97); 3 strains bad lower grepafloxacin MICs (2xc3x97); and one strain had lower sparfloxacin MICs (2xc3x97), suggesting that an efflux mechanism contributed to the raised MICs in some cases (Table 4).
Microbroth dilution MIC results of the 12 strains tested by time-kill are presented in Table 5. Microdilution MICs were all within one dilution of agar MICs. For the two quinolone resistant strains (both penicillin susceptible), gemifloxacin microbroth MICs were 0.5 and 0.25 xcexcg/ml, respectively. Time-kill results (Table 6) showed that levofloxacin at the MIC, gemifloxacin and sparfloxacin at 2xc3x97MIC and ciprofloxacin, grepafloxacin and trovafloxacin at 4xc3x97MIC, were bactericidal after 24 h. Various degrees of 90% and 99% killing by all quinolones was detected after 3 h. Gemifloxacin and trovafloxacin were both bactericidal at the microbroth MIC for the two quinolone resistant pneumococcal strains. Gemifloxacin was uniformly bactericidal after 24 h at xe2x89xa60.5 xcexcg/ml. Amoxicillin, at 2xc3x97MIC and cefuroxime at 4xc3x97MIC, were bactericidal after 24 h, with some degree of killing at earlier time periods. By contrast, macrolides gave slower killing against the 7 susceptible strains tested, with 99.9% killing of all strains at 2-4xc3x97MIC after 24 hours.
For the five quinolone susceptible strains tested for PAE, MICs were similar to those obtained by microdilution, with gemifloxacin having MICs of 0.25 xcexcg/ml against the quinolone resistant strain (MICs of other quinolones 4-32 xcexcg/ml). PAEs (h)(10xc3x97MIC) for the 5 quinolone susceptible strains ranged between 0.4-1.6 for gemifloxacin; 0.5-1.5 h for ciprofloxacin (except for the quinolone resistant strain which gave a ciprofloxacin PAE of 6.3); 0.9-2.3 (levofloxacin); 0.3-2.0 (sparfloxacin); 0.3-2.6 (grepafloxacin); 1.3-3.0 (trovafloxacin). At 5xc3x97MIC, PAEs (h) for the quinolone resistant strain were 0.9 (gemifloxacin); 3.7 (ciprofloxacin); 1.3 (levofloxacin); 1.5 (sparfloxacin); 1.5 (grepafloxacin); 1.3 (trovafloxacin). PAEs for non-quinolone compounds (10xc3x97MIC) ranged between 0.3-5.8 (amoxicillin); 0.8-2.9 (cefuroxime); 1.3-3.0 (azithromycin); 1.8-4.5 (clarithromycin).
A further embodiment of the present invention is based, in part, on experiments wherein in vitro activity of gemifloxacin was compared with that of ciprofloxacin, levofloxacin, sparfloxacin, grepafloxacin and trovafloxacin against 28 pneumococci with ciprofloxacin MICs xe2x89xa78 xcexcg/ml. Gemifloxacin MICs (xcexcg/ml) ranged between 0.03-1.0 (MIC50/90 0.25/0.5), compared with ciprofloxacin 8xe2x86x9232 (MIC50/90 16/ greater than 32), levofloxacin 4xe2x86x9232 (MIC50/90 16/ greater than 32), sparfloxacin 0.25xe2x86x9232 (MIC50/90 8/16), grepafloxacin 0.5-16 (MIC50/90 4/8) and trovafloxacin 0.25-8 (MIC50/90 1.0/4.0). DNA sequence analysis showed that all but one strain had a mutation in the quinolone resistance-determining region (QRDR) of parC and/or gyrA. Mutations in parC were at S79-F or Y, D83-G or N, N91-D, R95-C, or K137-N. Mutations in gyrA were at S81-A, C, F, or Y; E85-K; or S114-G. Twenty-one strains had a mutation in parE at D435-N or I460-V. Only two strains had a mutation in gyrB at D435-N or E474-K. Twenty-one strains had a total of three or four mutations in the QRDRs of parC, gyrA, pare, and gyrB. Of these 21 strains, all were resistant to ciprofloxacin (MIC xe2x89xa78 xcexcg/ml), levofloxacin (MIC xe2x89xa74 xcexcg/ml), and sparfloxacin (MIC xe2x89xa71 xcexcg/ml); 20 were resistant to grepafloxacin (MIC xe2x89xa71 xcexcg/ml) and 11 were resistant to trovafloxacin (MIC xe2x89xa72 xcexc/ml), yet gemifloxacin MICs were xe2x89xa60.5 xcexcg/ml in 19 of the strains. In the presence of reserpine, 23 strains had lower ciprofloxacin MICs (2-16xc3x97), 13 strains had lower gemifloxacin MICs (2-4xc3x97), 7 strains had lower levofloxacin MICs (2-4xc3x97); 3 strains had lower grepafloxacin MICs (2xc3x97) and one strain had lower sparfloxacin MICs (2xc3x97), indicating that an efflux mechanism contributed to the raised MICs in some cases. Results show that, irrespective of the mechanism of quinolone resistance, gemifloxacin showed the greatest in vitro activity against all pneumococcal strains tested. Against 28 strains with ciprofloxacin MICs xe2x89xa78 xcexcg/ml, gemifloxacin had the lowest MICs (0.03-1.0 xcexcg/ml, MIC90 0.5 xcexcg/ml), compared with MICs ranging between 0.25 to  greater than 32.0 xcexcg/ml (MIC90 s4.0xe2x86x9232.0 xcexcg/ml) for the other quinolones, with trovafloxacin, grepafloxacin, sparfloxacin and levofloxacin, in ascending order, giving the next lowest MICs (Table 7). Mechanisms of quinolone resistance are presented in Tables 8 and 9. As can be seen, quinolone resistance was associated with mutations in the quinolone resistance-determining region (QRDR) of parC, gyrA, parE and/or gyrB. Mutations in ParC were at S79-F or Y, D83-N, R95-C, or K137-N. Mutations in gyrA were at S83-A, C, F, or Y; E87-K; or S116-G. Twenty-one strains had a mutation in parE at D435-N or I460-V. Only two strains had a mutation in grB at D435-N or E474-K. Twenty-one strains had a total of three or four mutations in the QRDRs of parC, gyrA, pare and gyrB (Table 8). Amongst these 21 strains all were resistant to ciprofloxacin (MICs xe2x89xa78 xcexcg/ml), levofloxacin (MICs xe2x89xa74 xcexcg/ml), and sparfloxacin (MICs xe2x89xa71 xcexcg/ml), 20 were resistant to grepafloxacin (MICs xe2x89xa71 xcexcg/ml) and 11 were resistant to trovafloxacin (MICs xe2x89xa72 xcexcg/ml), yet gemifloxacin MICs were xe2x89xa60.5 xcexcg/ml in 19 of the strains (Table 8).
In the presence of reserpine 23 strains had lower ciprofloxacin MICs (2-16xc3x97), 13 strains had lower gemifloxacin MICs (2-4xc3x97), 7 strains bad lower levofloxacin MICs (2-4xc3x97); 3 strains had lower grepafloxacin MICs (2xc3x97); and one strain bad lower sparfloxacin MICs (2xc3x97), indicating that an efflux mechanism contributed to the raised MICs in some cases (Table 9). Previous studies have shown gemifloxacin to be 32 to 64 fold more active than ciprofloxacin, ofloxacin, sparfloxacin and trovafloxacin against methicillin-susceptible and -resistant Staphylococcus aureus, methicillin-resistant Ataphylococcus epidennidis and S. pneumoniae Gemifloxacin was also highly active against most members of the family Enterobacteriaceae, with activity which was more potent than those of sparfloxacin and ofloxacin and comparable to that of ciprofloxacin. Gemifloxacin was the most active agent against Gram positive species resistant to other quinolones and glycopeptides. Gemifloxacin has variable activity against anaerobes, and is very active against the Gram positive group (Cormican, et al., Antimicrobiol. Agents Chemother. 41 :204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; Oh, et al., Antimicrob. Agents Chenother. 40:1564-1568, 1996).
In our study, gemifloxacin gave significantly lower MICs against highly quinolone-resistant pneumococci, irrespective of quinolone resistance mechanism. This was the case in double mutants with mutations in both parC and gyrA, strains which have previously been shown to be highly resistant to other quinolones, as well as for strains with an efflux mechanism (Pan, et al., Antimicrob. Agents Chemother. 40:2321-2326, 1996 and Brenwald, et al., Antimicrob. Agents Chemother. 42:2032-2035, 1998).
In summary, gemifloxacin was the most potent quinolone tested against quinolone resistant pneumococci. The incidence of quinolone-resistant pneumococci is currently very low. However, this situation may change with the introduction of broad-spectrum quinolones into clinical practice, and in particular in the pediatric population, leading to selection of quinolone-resistant strains (Davies, et al., Antimicrob. Agents Chemother. 43:1177-1182, 1999). Results indicate that selective introduction of quinolones such as gemifloxacin into the pediatric environment is predicated upon toxicologic studies. Additionally, if the incidence of quinolone-resistant pneumococci increases, gemifloxacin will be a well-placed therapeutic option. Gemnifloxacin is a promising new antipneumococcal agent, irrespective of the strain""s susceptibility to quinolones and other agents.
II. Haemophilus Pathogens
Nine quinolone-resistant H. influenzae strains yielded MIC50s of 0.25 xcexcg/ml for gemifloxacin (highest MIC 1.0 xcexcg/ml) compared to 1.0 xcexcg/ml (highest MIC 4.0-8.0 xcexcg/ml) for the other quinolones tested (Table 10). Mechanisms of quinolone resistance in the H. influenzae strains are presented in Table 11. All nine strains had mutations at Ser 84 in GyrA with Ser 84 to Leu, Phe, or Tyr observed. Additional mutations in GyrA at Asp 88 to Asn or Tyr, and Ala 117 to Glu were also observed in some strains. Most strains also had at least one mutation in ParC (at Asp 83, Ser 84, Glu 88, Ser 133, or Asn 138) and ParE (at Gly 405, Asp 420, Ser 458, or Ser 474). Strain 4 had an in-frame insertion in parE that led to an insertion of a Ser residue in between Ser 458 and Thr 459. Only one strain had a mutation in GyrB (at Gln 468). The most resistant strain (strain 9) had double mutations in GyrA, ParC and ParE.
Previous studies have shown gemifloxacin to be 32-64 fold more active than ciprofloxacin, ofloxacin, sparfloxacin and trovafloxacin against methicillin-susceptible and -resistant S. aureus, methicillin-resistant Staphylococcus epideridis and S. pneumoniae Gemifloxacin was also highly active against most members of the family Enterobacteriaceae, with activity more potent than those of sparfloxacin and ofloxacin and comparable to that of ciprofloxacin. Gemifloxacin was the most active agent against Gram positive species resistant to other quinolones and glycopeptides. Gemifloxacin has variable activity against anaerobes and is very active against the Gram positive group (Cormican, et al., Antimicrob. Agents Chemother. 41:204-211, 1997; Hohl, et al., Clin. Microbiol. Infect. 4:280-284, 1998; and Oh, et al., Antimicrob. Agents Chemother. 40:1564-1568, 1996. In the studies set forth herein, only gemifloxacin gave MICs xe2x89xa61.0 xcexcg/ml against the rare strains of H. influenzae with raised quinolone MICs. Previous studies (Bootsma, et al., J. Antimicrob. Chemother. 39:292-293, 1997; Georgiou, et al., Antimicrob. Agents Chemother. 40:1741-1744, 1996; and Vila, et al., Antimicrob. Agents Chemother. 43:161-162, 1999) have shown that the primary target of quinolones in H. influenzae is GyrA; low-level resistance is associated with a mutation in GyrA (Ser 84 or Asp 88) and high-level resistance with an additional mutation in ParC (Asp 83, Ser 84 or Glu 88). Sequencing results from this study were in agreement with the above previous reports, as all nine strains had at least one mutation in GyrA and the most resistant strains (ciprofloxacin MICs xe2x89xa71.0 xcexcg/ml) had an additional mutation in ParC. Mutations were found in GyrA (Ala 117) and ParC (Ser 133, Asn 138) that have not been previously reported. Provided herein is a novel examination of mutations in GyrB and ParE in H. influenzae: most strains had mutations in ParE, but only one strain in GyrB. Of particular interest was insertion of a serine between serine 458 and threonine 459 of ParE in one strain. It, therefore, appears that ParE is more important in quinolone resistance in H. influenzae than GyrB.
Results of this study indicate excellent activity of gernifloxacin against quinolone-resistant i H. influenzae (including those with multiple mutations) by MIC. Because of the wide spectrum of activity of gemifloxacin against other respiratory pathogens, such as pneumococci (including quinolone-resistant strains), Legionella, mycoplasmas and chlamydia, this compound represents an attractive alternative to other quinolone and non-quinolone agents for empiric treatment of community-acquired respiratory tract infections.
All strains had mutations at position 84 in gyrA, and the most R strain had double mutations in gyrA, parC and parE. Strains with mutations at position 84 in parC and gyrA plus mutations in parE were tro R. Gem had the lowest MICs against all strains irrespective of their mutation mechanism.
The invention provides a method for modulating metabolism of a rare pathogenic H. influenzae strain. Skilled artisans can readily choose a rare pathogenic Haemophilus. influenzae strain or patients infected with or suspected to be infected with these organisms to practice the methods of the invention. Alternatively, the bacteria useful in the methods of the invention may be those described herein.
The invention provides a method for modulating metabolism of pneumococcal and Haemophilus pathogenic bacteria. Skilled artisans can readily choose pneumococcal and Haemophilus pathogenic bacteria or patients infected with or suspected to be infected with these organisms to practice the methods of the invention. Alternatively, the bacteria useful in the methods of the invention may be those described herein.
The contacting step in any of the methods of the invention may be performed in many ways that will be readily apparent to the skilled artisan. However, it is preferred that the contacting step is a provision of a composition comprising a gemifloxacin compound to a human patient in need of such composition or directly to bacteria in culture medium or buffer.
For example, when contacting a human patient or contacting said bacteria in a human patient or in vitro, the compositions comprising a quinolone, particularly a gemifloxacin compound, preferably pharmaceutical compositions may be administered in any effective, convenient manner including, for instance, administration by topical, oral, anal, vaginal, intravenous, intraperitoneal, intramuscular, subcutaneous, intranasal or intradermal routes among others.
It is also preferred that these compositions be employed in combination with a non-sterile or sterile carrier or carriers for use with cells, tissues or organisms, such as a pharmaceutical carrier suitable for administration to a subject. Such compositions comprise, for instance, a media additive or a therapeutically effective amount of a compound of the invention, a quinolone, preferably a gemifloxacin compound, and a pharmaceutically acceptable carrier or excipient. Such carriers may include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol and combinations thereof. The formulation should suit the mode of administration.
Quinolone compounds, particularly gemifloxacin compounds and compositions of the methods of the invention may be employed alone or in conjunction with other compounds, such as bacterial efflux pump inhibitor compounds or antibiotic compounds, particularly non-quinolone compounds, e.g., beta-actam antibiotic compounds.
In therapy or as a prophylactic, the active agent of a method of the invention is preferably administered to an individual as an injectable composition, for example as a sterile aqueous dispersion, preferably an isotonic one.
Alternatively, the gemifloxacin compounds or compositions in the methods of the invention may be formulated for topical application for example in the form of ointments, creams, lotions, eye ointments, eye drops, ear drops, mouthwash, impregnated dressings and sutures and aerosols, and may contain appropriate conventional additives, including, for example, preservatives, solvents to assist drug penetration, and emollients in ointments and creams. Such topical formulations may also contain compatible conventional carriers, for example cream or ointment bases, and ethanol or oleyl alcohol for lotions. Such carriers may constitute from about 1% to about 98% by weight of the formulation; more usually they will constitute up to about 80% by weight of the formulation.
For administration to mammals, and particularly humans, it is expected that the antibacterially effective amount is a daily dosage level of the active agent from 0.001 mg/kg to 10 mg/kg, typically around 0.1 mg/kg to 1 mg/kg, preferably about 1 mg/kg. A physician, in any event, will determine an actual dosage that is most suitable for an individual and will vary with the age, weight and response of the particular individual. The above dosages are exemplary of the average case. There can, of course, be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention. It is preferred that the dosage is selected to modulate metabolism of a bacteria in such a way as to inhibit or stop growth of said bacteria or by killing said bacteria. The skilled artisan may identify this amount as provided herein as well as using other methods known in the art, e.g. by the application MIC tests.
A further embodiment of the invention provides for the contacting step of the methods to further comprise contacting an in-dwelling device in a patient. In-dwelling devices include, but are not limited to, surgical implants, prosthetic devices and catheters, i.e., devices that are introduced to the body of an individual and remain in position for an extended time. Such devices include, for example, artificial joints, heart valves, pacemakers, vascular grafts, vascular catheters, cerebrospinal fluid shunts, urinary catheters, and continuous ambulatory peritoneal dialysis (CAPD) catheters.
A quinolone, particularly a gemifloxacin compound or composition of the invention may be administered by injection to achieve a systemic effect against relevant bacteria, preferably a pneumococcal or Haemophilus pathogenic bacteria, shortly before insertion of an in-dwelling device. Treatment may be continued after surgery during the in-body time of the device. In addition, the composition could also be used to broaden perioperative cover for any surgical technique to prevent bacterial wound infections caused by or related to pneumococcal or Haemophilus pathogenic bacteria.
In addition to the therapy described above, a gemifloxacin compound or composition used in the methods of this invention may be used generally as a wound treatment agent to prevent adhesion of bacteria to matrix proteins, particularly pneumococcal or Haemophilus pathogenic bacteria, exposed in wound tissue and for prophylactic use in dental treatment as an alternative to, or in conjunction with, antibiotic prophylaxis.
Alternatively, a quinolone, particularly a gemifloxacin compound or composition of the invention may be used to bathe an indwelling device immediately before insertion. The active agent will preferably be present at a concentration of 1 xcexcg/ml to 10 mg/ml for bathing of wounds or indwelling devices.
Also provided by the invention is a method of treating or preventing a bacterial infection by pneumococcal or Haemophilus pathogenic bacteria comprising the step of administering an antibacterially effective amount of a composition comprising a quinolone, particularly a gemifloxacin compound to a mammal, preferably a human, suspected of having or being at risk of having an infection with pneumococcal or Haemophilus pathogenic bacteria.
A preferred object of the invention provides a method wherein said pneumococcal pathogenic bacteria is selected from the group consisting of: bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parc, gyrA, parE, and/or gyrB; bacteria comprising a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; bacteria comprising a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; bacteria comprising a mutation in parE at D435-N or I460-V; bacteria comprising a mutation in gyrB at D435-N or E474-K; bacteria comprising at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in ParC at S79-F or Y, D83-N, R95-C, or K137-N; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in parE at D435-N or I460-V; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in gyrB at D435-N or E474-K; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; bacteria that are ciprofloxacin-resistant, levofloxacin-resistant, sparfloxacin-resistant, grepafloxacin-resistant, or trovafloxacin-resistant, or a combination thereof, that comprise a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB; Streptococcus pneumoniae bacteria comprising a mutation in ParC at 879-F or Y, D83-N, R95-C, or K137-N; Streptococcus pneumoniae bacteria comprising a mutation in gyrA at S83-A, C, F, or Y; E87-K; or S116-G; Streptococcus pneumoniae bacteria comprising a mutation in parE at D435-N or I460-V; Streptococcus pneumoniae bacteria comprising a mutation in gyrB at D435-N or E474-K; Streptococcus pneumoniae bacteria comprising at least four mutations in a QRDR or parC, gyrA, parE, and gyrB; and Streptococcus pneumoniae bacteria comprising a mutation in a quinolone resistance-determining region (QRDR) of parC, gyrA, parE, and/or gyrB.
A preferred object of the invention provides a method wherein said quinolone-resistant pneumococcal pathogenic bacteria is selected from the group consisting of: a pneumococcal strain comprising a mutation in the quinolone resitance-determining region (QRDR) of parC and/or gyrA; a pneumococcal strain comprising a mutation in parC said mutation comprising S79-F and/or Y, D83-G and/or N, N91-D, R95-C, and/or K137-N; a pneumococcal strain comprising a mutation in gyrA said mutation comprising S81-A, C, F, and/or Y; E85-K; and/or S114-G; a pneumococcal strain comprising a mutation in parE said mutation comprising D435-N and/or I460-V; a pneumococcal strain comprising a mutation in gyrB said mutation comprising D435-N and/or E474-K; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB; a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB, any of which are resistant to ciprofloxacin, levofloxacin, or sparfloxacin; and a pneumococcal strain comprising a mutation in comprising three or four mutations in a QRDRs of parC, gyrA, parE, and/or gyrB, any of which also comprising an efflux mechanism of quinolone resistance.
A further preferred object of the invention provides a method wherein said rare pathogenic H. influenzae strain is selected from the group consisting of: bacteria comprising a mutation set forth in Table 11 or 12; a Haemophilus influenzae strain set forth in Table 11 or 12; bacteria of the genus Haemophilhs comprising a mutation set forth in Table 11 or 12; and bacteria of the species Haemophilus influenzae comprising a mutation set forth in Table 11 or 12.
Other pneumococcal and Haemophilus pathogenic bacteria may also be included in the methods. The skilled artisan may identify these organisms as provided herein as well as using other methods known in the art, e.g. MIC tests.
Preferred embodiments of the invention include, among other things, methods wherein said composition comprises gemifloxacin, or a pharmaceutically acceptable derivative thereof.